Application of long-chain non-coding RNA in preparation of hepatocellular carcinoma diagnostic product or therapeutic drug

A technology of long-chain non-coding and hepatocellular carcinoma, which is applied in the application field of long-chain non-coding RNA in the preparation of diagnostic products or therapeutic drugs for hepatocellular carcinoma. It can solve problems such as functions that have not been reported, and improve the level of targeted therapy , the effect of promoting proliferation

Active Publication Date: 2021-03-09
THE NAVAL MEDICAL UNIV OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The function of lncRNA-RNA pol I regulated has not been reported yet

Method used

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  • Application of long-chain non-coding RNA in preparation of hepatocellular carcinoma diagnostic product or therapeutic drug
  • Application of long-chain non-coding RNA in preparation of hepatocellular carcinoma diagnostic product or therapeutic drug
  • Application of long-chain non-coding RNA in preparation of hepatocellular carcinoma diagnostic product or therapeutic drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0039] The present invention collects liver cancer cell line Huh7 and a certain amount of hepatocellular carcinoma and paracancerous tissue samples, extracts RNA and reverse transcribes it into cDNA, and uses Real-time PCR method to analyze the expression level of lncRNA-RNA pol I regulated, the specific method is as follows :

[0040] 1. Collection of samples and RNA extraction:

[0041] (1) Sample collection

[0042] a. Tissue samples were collected at the sample bank of Shanghai Oriental Hepatobiliary Hospital. Snip an appropriate amount of hepatocellular carcinoma tissue, paracancerous liver tissue (within 2cm from the border) and liver tissue away from the cancer border (more than 2cm from the border), wash with PBS three times, put into a centrifuge tube containing 1ml Trizol, and use Homogenize the tissue with a homogenizer and place it on ice for 15 minutes.

[0043] b. The cell samples are kept by the experimental center. The cells cultured in the 6-well plate wer...

Embodiment 2

[0061] 1. Design of siRNA (small interfering RNA): Through BLAST retrieval, design three pairs of siRNA in the specific sequence region of lncRNA-RNA pol Iregulated, as shown in Table 1:

[0062] Table 1: Sequences of siRNAs

[0063]

[0064] The above siRNA is added at the 3' end TT To enhance the stability of siRNA.

[0065] It was confirmed by RT-PCR that the siRNA was transfected into Huh7 cells (purchased from the Cell Bank of Shanghai Chinese Academy of Sciences). Transfection method: take the logarithmically grown cells, inoculate them on a culture plate, add culture medium, and cultivate for 48 hours (cells grow to 70-90% full), perform transfection experiments according to the instructions of lipofectamine2000 (invitrogen), and collect RNA after 24 hours with Real -time PCR was used to detect the expression of lncRNA-RNA pol I regulated.

[0066] Results: Compared with the negative control group, the expression of lncRNA-RNA pol Iregulated in the siRNA1, siRNA2...

Embodiment 3

[0067] Example 3: Effect of lncRNA-RNA pol I regulated on the proliferation ability of liver cancer cells

[0068] Huh7 cells were inoculated in a 96-well cell culture plate with a cell aliquoter, 3000 cells / well. After 18 hours, lncRNA-RNA pol I regulated specific siRNA and negative control siRNA were transfected, with 3 replicates in each group. The dosage of Lipofectamine2000 was 0.25 μl / well, and the dosage of RNA was 5 pmol / well. After 6 hours, the medium was changed, and the culture was continued. After 54 hours of transfection, the medium was aspirated, and each well was replaced with DMEM complete medium containing 10% CCK8 to continue culturing for 1 hour. The absorbance at 450 nm was detected on a microplate reader. It was found that the OD value of the lncRNA-RNA pol I regulated specific siRNA interference group was significantly less than that of the control group ( Figure 4 ). The above results indicate that lncRNA-RNA pol I regulated can significantly promote...

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Abstract

The invention relates to the field of gene therapy and medical diagnosis, and particularly relates to application of long-chain non-coding RNA in preparation of a hepatocellular carcinoma diagnostic reagent or kit. The invention also provides application of the long-chain non-coding RNA in preparation of a hepatocellular carcinoma therapeutic drug. The invention finds that lncRNA-RNA Pol I regulated is a long-chain non-coding RNA highly expressed in hepatocellular carcinoma tissues for the first time, and provides a new diagnostic marker and therapeutic target for hepatocellular carcinoma.

Description

technical field [0001] The invention relates to the fields of gene therapy and medical diagnosis, in particular, the application of a long-chain non-coding RNA in the preparation of diagnostic products or therapeutic drugs for hepatocellular carcinoma. Background technique [0002] Primary hepatocellular carcinoma (hereinafter referred to as liver cancer) is one of the common malignant tumors in my country. Due to the limited means of early diagnosis and treatment, the death cases and diagnosed cases of liver cancer are almost equal every year. Therefore, exploring the relationship between the function of new genes and the occurrence and development of liver cancer is of great significance for revealing the molecular mechanism of liver cancer, designing therapeutic drugs and judging the prognosis, and further improving the diagnosis and treatment of liver cancer in my country. [0003] Long non-coding RNA (long non-coding RNA, lncRNA) is a class of RNA molecules with transc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886A61K45/00A61K31/713A61P1/16A61P35/00
CPCC12Q1/6886A61K45/00A61K31/713A61P1/16A61P35/00C12Q2600/158C12Q2600/178C12Q2600/118C12Q2600/136
Inventor 杨富戴智慧葛春梅王月帆尹昊瓒
Owner THE NAVAL MEDICAL UNIV OF PLA
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